1. Field of the Invention
The present invention relates to a novel DNA and a gene comprising thereof, the DNA having Plexin-like sequence, being expressed specifically in vessels throughout all angiogenesis period from the beginning of angiogenesis in ontogenesis, being intensely expressed in stratified squamous epithelium in stomach of grown-up adults, being moderately expressed in uterus, and being expressed in various organs including brain; a novel polypeptide encoded by the DNA and a recombinant protein comprising the polypeptide; an antibody against the novel polypeptide; a screening method of an in vivo angiogenesis proliferation/differentiation control factor and compound which controls vascular cells proliferation/differentiation and is involved in inhibition of vascular proliferation which supports proliferation of cancer cells; and a measurement kit for the vascular proliferation/differentiation control activity of the differentiation control factor and compound.
2. Description of the Related Art
As the human genome project and the human cDNA project have successfully completed, many disease-related genes or candidates thereof have been identified. However, researches using human genes are limited due to ethical constraints, so that novel approach is being explored. In this circumstance, identification of homologous genes in model organisms is a crucial step to facilitate the research. Rodents, particularly mouse is a model organism which has been studied in depth so far. Relatively abundant data on genome or mutants of the mouse is available, but it is not still sufficient yet. On the other hand, as a means to analyze genes which are expressed in vivo, researches to analyze cDNA sequences randomly has been conducted, and the sequences of cDNA fragments identified in the researches have been registered as Expressed Sequence Tag (EST) in databases for publication. However, many EST has only limited data for base sequences of 100 bps to 500 bps in length. It is difficult to estimate the function of a base sequence from the data thus accumulated.
In various organs, physiological functions are regulated under control by many hormones, hormone-like substances, neurotransmitters, and physiologically active substances. Also, in the regulation of the physiological functions of various organs, proliferation, guidance and activation of specific cells responsible for the function is involved. Therefore, in order to develop a new drug, it is useful to obtain a novel gene which is specifically expressed in various organs, such as a gene expressed specifically in vessels throughout all angiogenesis period from the beginning of angiogenesis in ontogenesis, and to generate a protein which is encoded by this gene which regulates complicated functions in angiogenesis and various organs. Furthermore, in order to efficiently screen agonists and antagonists of a protein for the purpose of drug development, it is necessary to estimate the function of gene encoding the protein expressed in vivo using homology search, to generate a recombinant protein based on the information by expressing the gene in appropriate expression system, and to generate antibodies which specifically bind to the protein. In an experimental system which models angiogenesis using mouse ES cells, attempts have been made to comprehensively obtain genes involved in angiogenesis, and basic information is now being accumulated (document 14). However, a gene which is involved in angiogenesis and has 4000 or more bps, has not been reported yet from rodents especially from a mouse.
In the Long Chain Human cDNA project, human KIAA0620 gene derived from human brain was reported (Ishikawa K., Nagase T., Suyama M., Miyajima N., Tanaka A., Kotani H., Nomura N. and Ohara, O., DNA Res., 1998, 5:169-176, Prediction of the coding sequences of unidentified human genes X. The complete sequences of 100 new cDNA clones from a brain which can code for large proteins in vitro., GenBank Accession No. AB014520, 6,754 bp, 1746aa, Homosapiens, cDNA, KIAA0620, Ohara O. et al.). Also it is reported that mRNA of a gene relating to human KIAA0620 is expressed in vascular endothelial cells or central nervous system (CNA) during development processes of a mouse (van der Zwaag, B. et al., Dev. Dyn., 2002, 225:336-343). But the function of the gene has not been identified.
Human KIAA0620 gene is located on the chromosome 3q21.3, and several SNPs of the human KIAA0620 gene are reported. As majority of human diseases are not caused by mere deletion of a specific gene, but by partial alteration in functions or activities of a protein by amino acid substitution, it is suspected that human KIAA0620 gene may be involved in healing of wound, healing of fracture, vascular occlusion and collateral vessel formation, periodic formation of vascular network in tunica mucosa uteri (transient or at the time of luteinization); various processes in which angiogenesis is undesirably involved such as proliferation of cancer cells, chronic articular rheumatism, diabetic retinopathy, endometriosis, obesity; and various processes in which angiogenesis is desirably involved such as heart attack, neurodegenerative diseases, circulatory deficits in legs, arteriosclerosis obliterans, and psoriasis vulgaris. Although it was reported that mRNA of the gene relating to human KIAA0620 gene is expressed in vascular endothelial cells or central nervous system (CNS) in a mouse during development process, researches using not human but experimental animals are essential to elucidate human KIAA0602 gene and its relationship. However, the gene that corresponds to human KIAA0620 gene has not been obtained yet from rodents particularly from a mouse which is an important model animal in researches of human pathology, thus a study that uses the gene could not have been conducted so far.    Non-Patent Document 1:1    Masashi Shibuya, 1999, Experimental Medicine, 17:712-715, Today's studies about angiogenesis and molecular regulatory mechanism    Non-Patent Document 2:2    Masanori Hirashima, Shinichi Nishikawa, 1999, Experimental Medicine, 17:716-720, Embryology of vascular endothelial cells    Non-Patent Document 3:3    Jun Yamashita, 2001, Experimental Medicine, 19:830-835, Angiogensis from embryonic stem cells    Non-Patent Document 4:4    Nobuyuki Takakura, 2001, Experimental Medicine, 19:836-840, Vascular cells and angiogenesis    Non-Patent Document 5:5    Chisa Sunami, Hiroyuki Shibata, Yuji Seki, 2001, Experimental Medicine, 19:841-846, Formation of cartilage and bone, and vascular invasion    Non-Patent Document 6:6    Nariaki Matsuura, Yoshihisa Okazaki, Naoyuki Tani, Hidetoshi Eguchi, 1999, Experimental Medicine, 17:741-752, Angiogenesis inhibitors produced by tumor    Non-Patent Document 7:7    Jane R. K. & Karmeri P. F., 2002, Nikkei Science, March issue, 22-29, Angiogenesis—new field of medicine    Non-Patent Document 8:8 van der Zwaag, B. et al., Dev. Dyn., 2002, 225:336-343, PLEXIN-D1, a novel plexin family member, is expressed in vascular endothelium and central nervous system during mouse embryogenesis.    Non-Patent Document 9:9    Tamagnone L, Artigiani S, Chen H, He Z, Ming G I, Song H, Chedotal A, Winberg M L, Goodman C S, Poo M, Tessier-Lavigne M, Comoglio P M., 2001, Cell, 99:71-80, Plexins are a large family of receptors for transmembrane, secreted, and GPI-anchored semaphorins in vertebrates    Non-Patent Document 10:10    Manahan, D., 1997, Science, 277:48-50, Signaling vascular morphogenesis and maintenance    Non-Patent Document 11:11    Barinaga, M., 1997, Science, 275:482-484, Designing therapies that target tumor blood vessels    Non-Patent Document 12:12    Asahara T, Murohara T, Sullivan A, Silver M, van der Zee R, Li T, Witzenbichler B, Schatteman G, Isner J M. Asahara, T. et al., 1997, Science, 275:964-967, Isolation of putative progenitor endothelial cells for angiogenesis    Non-Patent Document 13:13    Risau, W., 1997, Nature, 386:671-674, Mechanisms of angiogenesis    Non-Patent Document 14:14    Yamashita J, Itoh H, Hirashima M, Ogawa M, Nishikawa S, Yurugi T, Naito M, Nakao K, Nishikawa S., Flk1-positive cells derived from embryonic stem cells serve as vascular progenitors    Non-Patent Document 15:15    Ishikawa K, Nagase T, Suyama M, Miyajima N, Tanaka A, Kotani H, Nomura N, Ohara O., 1998, DNA Res., 5:169-76, Prediction of the coding sequences of unidentified human genes X. The complete sequences of 100 new cDNA clones from brain which can code for large proteins in vitro    Non-Patent Document 16:16    Tamagnone L, & Comoglio P M., 2000, Trends in Cell Biology, 10:377-383, Signalling by semaphorin receptors: cell guidance and beyond    Non-Patent Document 17:17    Shimizu M, Murakami Y, Suto F, Fujisawa H, 2000, J. Cell Biol., 148:1283-1293, Determination of cell adhesion sites of neuropilin-1